Whole-Genome Sequencing: Automated, Nonindexed Library Preparation

descriptionPublicationkeyboard_double_arrow_right Article 01 Nov 2016 English Publisher:Cold Spring Harbor LaboratoryJournal:Cold Spring Harbor Protocols, volume 2,017, page pdb.prot094623 (issn: 1940-3402, eissn: 1559-6095,

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Authors: Elaine, Mardis; W Richard, McCombie;

doi: 10.1101/pdb.prot094623 , 10.1101/pdb.prot094649

pmid: 27803279 , 27803270

Whole-Genome Sequencing: Automated, Nonindexed Library Preparation

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Abstract

This protocol describes an automated procedure for constructing a nonindexed Illumina DNA library and relies on the use of a CyBi-SELMA automated pipetting machine, the Covaris E210 shearing instrument, and the epMotion 5075. With this method, genomic DNA fragments are produced by sonication, using high-frequency acoustic energy to shear DNA. Here, double-stranded DNA is fragmented when exposed to the energy of adaptive focused acoustic shearing (AFA). The resulting DNA fragments are ligated to adaptors, amplified by polymerase chain reaction (PCR), and subjected to size selection using magnetic beads. The product is suitable for use as template in whole-genome sequencing.

Keywords

Automation, Laboratory, Sonication, DNA Ligases, DNA, Genomics, Sequence Analysis, DNA, Molecular Biology, Polymerase Chain Reaction, Gene Library

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